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Background of 14-3-3 protein epsilon antibody

The 14-3-3 family of proteins plays a key regulatory role in sigl transduction, checkpoint control, apoptotic and nutrient-sensing pathways. 14-3-3 proteins are highly conserved and ubiquitously expressed. There are at least seven isoforms, ß, ?, e, s, ?, t, and ? that have been identified in mammals. The initially described a and d isoforms are confirmed to be phosphorylated forms of ß and ?, respectively. Through their amino-termil a helical region, 14-3-3 proteins form homo- or heterodimers that interact with a wide variety of proteins: transcription factors, metabolic enzymes, cytoskeletal proteins, kises, phosphatases, and other sigling molecules. The interaction of 14-3-3 proteins with their targets is primarily through a phospho-Ser/Thr motif. However, binding to divergent phospho-Ser/Thr motifs, as well as phosphorylation-independent interactions, has been observed. 14-3-3 binding masks specific sequences of the target protein and therefore modulates target protein localization, phosphorylation state, stability, and molecular interactions. 14-3-3 proteins may also induce target protein conformatiol changes that modify target protein function. Distinct temporal and spatial expression patterns of 14-3-3 isoforms have been observed in development and in acute response to extracellular sigls and drugs, suggesting that 14-3-3 isoforms may perform different functions despite their sequence similarities. Several studies suggest that 14-3-3 isoforms are differentially regulated in cancer and neurological syndromes.

The extracts of mouse brain (50ug) were resolved by SDS-PAGE, transferred to NC membrane and probed with anti-human 14-3-3 epsilon (1:1000). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system.

Western blot analysis: The extracts of mouse brain (50 ug) were resolved by SDS-PAGE, transferred to NC membrane and probed with anti-human 14-3-3 epsilon (1:1000). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system.

Immunofluorescent analysis of 14-3-3 epsilon staining in NIH3T3 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).

Predicted cell location: Cytoplasm. Positive control: Human colon cancer tissue. Recommended dilution: 1/50-200 The image on the left is immunohistochemistry of paraffin-embedded Human colon cancer tissue using YWHAE antibody at dilution 1/60, on the right is treated with the synthetic peptide. (Original magnification:x200)